Fernández-Nestosa María J, Guimerà Nuria, Sanchez Diego F, Cañete-Portillo Sofía, Velazquez Elsa F, Jenkins David, Quint Wim, Cubilla Antonio L
*Polytechnic School §School of Medicine, National University of Asunción †Instituto de Patología e Investigación, Asunción, Paraguay ‡DDL Diagnostic Laboratory, Rijswijk, The Netherlands ∥Miraca Life Sciences and Tufts University, Boston, MA.
Am J Surg Pathol. 2017 Jun;41(6):820-832. doi: 10.1097/PAS.0000000000000821.
Laser capture microdissection-polymerase chain reaction (LCM-PCR) supported by p16 was used for the first time to demonstrate human papillomavirus (HPV) DNA in histologically specific penile lesions, which were as follows: squamous hyperplasia (12 lesions, 10 patients), flat lesions (12 lesions, 5 patients), condylomas (26 lesions, 7 patients), penile intraepithelial neoplasia (PeIN) (115 lesions, 43 patients), and invasive squamous cell carcinomas (26 lesions, 26 patients). HPV was detected by whole-tissue section and LCM-PCR. LCM proved to be more precise than whole-tissue section in assigning individual genotypes to specific lesions. HPV was negative or very infrequent in squamous hyperplasia, differentiated PeIN, and low-grade keratinizing variants of carcinomas. HPV was strongly associated with condylomas, warty/basaloid PeIN, adjacent flat lesions, and warty/basaloid carcinomas. A single HPV genotype was found in each lesion. Some condylomas and flat lesions, especially those with atypia, were preferentially associated with high-risk HPV. Unlike invasive carcinoma, in which few genotypes of HPV were involved, there were 18 HPV genotypes in PeIN, usually HPV 16 in basaloid PeIN but marked HPV heterogeneity in warty PeIN (11 different genotypes). Variable and multiple HPV genotypes were found in multicentric PeIN, whereas unicentric PeIN was usually related to a single genotype. There was a correspondence among HPV genotypes in invasive and associated PeIN. p16 was positive in the majority of HPV-positive lesions except condylomas containing LR-HPV. p16 was usually negative in squamous hyperplasia, differentiated PeIN, and low-grade keratinizing variants of squamous cell carcinomas. In summary, we demonstrated that LCM-PCR was a superior research technique for investigating HPV genotypes in intraepithelial lesions. A significant finding was the heterogeneity of HPV genotypes in PeIN and the differential association of HPV genotypes with subtypes of PeIN. The presence of atypia and high-risk HPV in condylomas and adjacent flat lesions suggests a precursor role, and the correspondence of HPV genotypes in invasive carcinomas and associated PeIN indicates a causal relation. Data presented support the bimodal hypothesis of penile cancer carcinogenesis in HPV-driven and non-HPV-driven carcinomas and justify the current WHO pathologic classification of PeIN in special subtypes.
首次使用由p16支持的激光捕获显微切割-聚合酶链反应（LCM-PCR）来检测组织学上特定的阴茎病变中的人乳头瘤病毒（HPV）DNA，这些病变如下：鳞状上皮增生（12个病变，10例患者）、扁平病变（12个病变，5例患者）、尖锐湿疣（26个病变，7例患者）、阴茎上皮内瘤变（PeIN）（115个病变，43例患者）以及浸润性鳞状细胞癌（26个病变，26例患者）。通过全组织切片和LCM-PCR检测HPV。在将个体基因型分配到特定病变方面，LCM被证明比全组织切片更精确。HPV在鳞状上皮增生、分化型PeIN和低级别角化型癌中呈阴性或非常罕见。HPV与尖锐湿疣、疣状/基底样PeIN、相邻扁平病变以及疣状/基底样癌密切相关。每个病变中仅发现一种HPV基因型。一些尖锐湿疣和扁平病变，尤其是那些具有异型性的病变，优先与高危型HPV相关。与浸润性癌不同，浸润性癌中涉及的HPV基因型较少，而PeIN中有18种HPV基因型，基底样PeIN中通常为HPV 16，但疣状PeIN中HPV具有明显的异质性（11种不同基因型）。在多中心PeIN中发现了可变和多种HPV基因型，而单中心PeIN通常与单一基因型相关。浸润性癌和相关PeIN中的HPV基因型之间存在对应关系。除了含有低风险HPV的尖锐湿疣外，大多数HPV阳性病变中的p16呈阳性。p16在鳞状上皮增生、分化型PeIN和鳞状细胞癌的低级别角化型变体中通常为阴性。总之，我们证明LCM-PCR是研究上皮内病变中HPV基因型的一种优越的研究技术。一个重要发现是PeIN中HPV基因型的异质性以及HPV基因型与PeIN亚型的差异关联。尖锐湿疣和相邻扁平病变中异型性和高危型HPV的存在提示其前驱作用，浸润性癌和相关PeIN中HPV基因型的对应关系表明存在因果关系。所呈现的数据支持阴茎癌致癌作用的双峰假说，即HPV驱动型和非HPV驱动型癌，并证明了世界卫生组织目前对特殊亚型PeIN的病理分类是合理的。
